Hamdy Mohammed El Sayed, Elsadek Fakhr Ahmed, Mohammed El Sayed Hanan, Al Johery Said Abd Elmohsen, Abdel Ghani Hassanein Wesam
Department of Botany, Faculty of Science, Zagazig University, Zagazig 44511, Egypt.
Department of Medical Microbiology & Immunology, Faculty of Medicine, Zagazig University, Zagazig 44511, Egypt.
Int J Microbiol. 2016;2016:8382605. doi: 10.1155/2016/8382605. Epub 2016 Mar 31.
Carbapenem-resistant Gram-negative bacilli resulting from β-lactamases have been reported to be an important cause of nosocomial infections and are a critical therapeutic problem worldwide. This study aimed to describe the prevalence of imipenem-resistant Gram-negative bacilli isolates and detection of bla VIM, bla TEM, bla SHV, bla CTX-M-1, and bla CTX-M-9 genes in these clinical isolates in Egyptian hospitals. The isolates were collected from various clinical samples, identified by conventional methods and confirmed by API 20E. Antibiotic susceptibility testing was determined by Kirby-Bauer technique and interpreted according to CLSI. Production of bla VIM, bla TEM, bla SHV, and bla CTX-M genes was done by polymerase chain reaction (PCR). Direct sequencing from PCR products was subsequently carried out to identify and confirm these β-lactamases genes. Out of 65 isolates, (46.1%) Escherichia coli, (26.2%) Klebsiella pneumoniae, and (10.7%) Pseudomonas aeruginosa were identified as the commonest Gram-negative bacilli. 33(50.8%) were imipenem-resistant isolates. 22 isolates (66.7%) carried bla VIM, 24(72.7%) had bla TEM, and 5(15%) showed bla SHV, while 12(36%), 6(18.2%), and 0(0.00%) harbored bla CTX-M-1, bla CTX-M-9, and bla CTX-M-8/25, respectively. There is a high occurrence of β-lactamase genes in clinical isolates and sequence analysis of amplified genes showed differences between multiple SNPs (single nucleotide polymorphism) sites in the same gene among local isolates in relation to published sequences.
据报道,由β-内酰胺酶引起的耐碳青霉烯类革兰氏阴性杆菌是医院感染的重要原因,也是全球关键的治疗难题。本研究旨在描述埃及医院临床分离株中耐亚胺培南革兰氏阴性杆菌的流行情况,并检测这些临床分离株中bla VIM、bla TEM、bla SHV、bla CTX-M-1和bla CTX-M-9基因。从各种临床样本中收集分离株,通过常规方法进行鉴定,并经API 20E确认。采用 Kirby-Bauer 技术进行抗生素敏感性试验,并根据CLSI进行解释。通过聚合酶链反应(PCR)检测bla VIM、bla TEM、bla SHV和bla CTX-M基因的产生。随后对PCR产物进行直接测序,以鉴定和确认这些β-内酰胺酶基因。在65株分离株中,最常见的革兰氏阴性杆菌为大肠埃希菌(46.1%)、肺炎克雷伯菌(26.2%)和铜绿假单胞菌(10.7%)。33株(50.8%)为耐亚胺培南分离株。22株(66.7%)携带bla VIM,24株(72.7%)有bla TEM,5株(15%)显示bla SHV,而分别有12株(36%)、6株(18.2%)和0株(0.00%)携带bla CTX-M-1、bla CTX-M-9和bla CTX-M-8/25。临床分离株中β-内酰胺酶基因的发生率较高,扩增基因的序列分析显示,与已发表序列相比,本地分离株中同一基因的多个单核苷酸多态性(SNP)位点存在差异。